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Cells are important in the study of virus isolation and identification, viral pathogenic mechanisms and antiviral immunity. The spotted knifejaw (Oplegnathus punctatus) is a significant farmed fish in China that has been greatly affected by diseases in recent years. In this study, a new cell line derived from the spotted knifejaw brain (SKB) was established and characterized. SKB cells multiplied well in Leibovitz's L‐15 medium supplemented with 10% fetal bovine serum at 28°C. Chromosome analysis revealed that modal chromosome number was 48 for SKB. SKB cells exhibit susceptibility to several fish viruses, such as a largemouth bass virus, red grouper nervous necrosis virus (RGNNV), infectious spleen and kidney necrosis virus (ISKNV), Singapore grouper iridovirus (SGIV) and spotted knifejaw iridovirus isolate (SKIV‐TJ), as shown by cytopathic effect and increased viral titers. Electron microscopy results showed that the cytoplasm contained a large number of vacuoles, and many virus particles existed at the edge of the vacuoles in RGNNV‐infected cells and numerous viral particles were scattered throughout the cytoplasm in both ISKNV‐ and SKIV‐TJ‐infected cells. These results suggest that SKB is an ideal tool for studying host–virus interactions and potential vaccine development. [ABSTRACT FROM AUTHOR]

Chinese perch (Siniperca chuatsi), an important fish for the aquaculture industry of China, is often affected by viral diseases. A stable and sensitive cell line can play an important role in virus identification and isolation, functional gene identification, virus pathogenic mechanism and antiviral immunity study. In the present study, a new cell line (S. chuatsi skin cell, SCSC) derived from the skin of S. chuatsi was established. The SCSC mainly consisted of fibroblastic‐like cells, which grew well in M199 medium supplemented with 10% foetal bovine serum at 25°C. Chromosome analysis revealed that the SCSC (44%) has a diploid chromosome number of 2n = 48. The SCSC can be transfected and expressed exogenous gene efficiently. It also showed high sensitivity to several aquatic animal viruses from different families including Rhabdoviridae, Iridoviridae and Reoviridae. In addition, RT‐PCR showed that S. chuatsi rhabdovirus (SCRV) started genome replication as early as 3 h post infection in the cells, which also induced the up‐regulation of a variety of immune‐related genes including these related to interleukin family, pattern recognition receptors, JAK–STAT pathway and interferon regulatory factors. In summary, current study provided a new tool in research of fish viruses and its interaction with host. [ABSTRACT FROM AUTHOR]

Aeromonas veronii is a freshwater bacterium associated with many diseases in aquatic animals. In this study, the pathogenic of A. veronii was isolated from diseased Macrobrachium nipponense showing signs of weakness, poor appetite and red gills in some farms in Jiangsu province, China. The pathogenicity of A. veronii to M. nipponense was determined by challenge experiments. Results of artificial challenge showed A. veronii NJ1 was pathogenic to M. nipponense, and the LD50 was 4.5 × 106 CFU/ml at 7 d post infection. Detection virulence factors of the A. veronii NJ1 showed that A. veronii NJ1 produced caseinase, lipase, amylase, lecithinase, gelatinase and haemolysin. Detection of virulence‐related genes showed that A. veronii NJ1 was positive for aerolysin (aer), outer membrane protein (ompA), adhesin (aha), flagellin (flaA, flgH, flgM, flgN), lipoidase (lip), cytotonic enterotoxins (act, alt, ast), serine protease (ser), DNases (exu), haemolysin (hlyA), type IV pilus (tapA) and fimbrillin (fim). Antibiotic sensitivity testing indicated A. veronii NJ1 was susceptible to quinolones, but resistant to penicillins. Additionally, the immune‐related gene expression in M. nipponense was evaluated, and the results showed that anti‐lipopolysaccharide factor (ALF1, ALF2, ALF3), Cusrin1, Cusrin2, Cusrin3, Haemocyanin, i‐lysozyme and Prophenoloxidase were significantly upregulated from 6 to 24 hr after A. veronii infection. The results of our study suggested that A. veronii was an aetiological element in the mass mortalities of M. nipponense and this study provided preliminary insights into the diversity in the immune response of M. nipponense to the A. veronii infection. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is one of the most prevalent neoplasms worldwide, particularly in China. Immune-related genes (IRGs) and immune infiltrating lymphocytes play specific roles in tumor growth. Considering how important immunotherapy has become for HCC treatment in the past decade, our objective was to establish a prognostic model by screening survival-related IRGs in patients with HCC. Using edgeR , we identified differentially expressed IRGs (DEIRGs), DEmiRNAs, and DElncRNAs. Functional enrichment analysis of DEIRGs was performed to investigate the biological functions of IRGs via gene ontology annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Protein-protein interaction and competing endogenous RNA networks were established using Cytoscape. Survival-associated IRGs were selected via univariate COX regression analysis, a The Cancer Genome Atlas (TCGA) prognostic model and GSE76427 validation model were developed using multivariate COX regression analysis test by AIC (Akaike Information Criterion). We identified 116 DEIRGs in patients with HCC; the "cytokine-cytokine receptor interaction" pathway was found to be the most enriched pathway. Via the prognostic model helped us classify patients into high- and low-risk score groups based on overall survival (OS); high risk score was associated with worse OS, and a positive correlation was observed between the prognostic model and immune cell infiltration. To summarize, we established a prognostic model using survival-related IRGs that provides sufficient information for prognosis prediction and immunotherapy of patients with HCC. [ABSTRACT FROM AUTHOR]

Ginkgo biloba leaf is widely used in traditional medicine in China. The present study aimed to illustrate the effects of dietary Ginkgo biloba leaf extract (GBLE) on growth performance and immune responses in common carp infected by Aeromonas hydrophila. Six different diets either not treated (control) or treated with 0.5, 1, 2, 5 and 10 g/kg of GBLE were designed to feed the fishes for 8 weeks. The results indicated that, compared to the control groups, 10 g/kg dietary GBLE significantly increased body growth and feed utilization. In GBLE dietary groups, red blood cell levels, white blood cells, hematocrit, hemoglobin, total protein, albumin and globulin were significantly increased relative to the control groups. Dietary supplementation with 5 g/kg GBLE increased the phagocytic ratio, and phagocytic indexes increased in the 2, 5 and 10 g/kg groups relative to the control groups. Moreover, 2, 5 and 10 g/kg GBLE diets increased O 2 − production compared to the control groups. Additionally, GBLE diets stimulated lysozyme activity (in 10 g/kg group) and inhibited bactericidal activity (in 0.5, 2, 5 and 10 g/kg group). Quantitative real-time PCR showed that IL1β, IL8, TNF-α, IL10, TGFβ, and inducible enzyme genes were prone to decrease while SAA, hepcidin and GPX1 were increased due to the GBLE diet in the intestine. In the head-kidney, the GBLE treatment decreased IL1β, IL8, TNF-α, IL10, TGFβ, INOS and arginase gene expressions, whereas SOD upregulation was found in the GBLE condition. The mRNA expressions of IL1β, IL8, TNF-α, IL10 and INOS were decreased, but SAA, hepcidin, GPX1 and SOD mRNA levels were increased in the spleen in the GBLE diet compared to the control. Additionally, diet supplemented with GBLE improved the survival rate infected with A. hydrophila. Our observations suggest that GBLE effectively enhanced growth performance, modulated immune-related gene expression. It improved survival rate of common carp after A. hydrophila infection and the optimum concentration we recommend is 10 g/kg of GBLE. • Dietary 10 g/kg GBLE significantly increased body growth and feed utilization in common carp. • Dietary GBLE stimulated immunity of common carp. • Dietary GBLE affected crucial immune gene expression in common carp. • Diet supplemented with GBLE improved the survival rate of common carp. [ABSTRACT FROM AUTHOR]

Clade 2.3.4.4 H5 avian influenza viruses (AIVs) are widely prevalent and of significant concern to the poultry industry and public health in China. Nowadays, the clade 2.3.4.4 H5N6 virus has become a dominant AIV subtype among domestic ducks in southern China. We found that waterfowl-origin clade 2.3.4.4 H5N6 viruses (A/goose/Guangdong/16568/2016, GS16568 and A/duck/Guangdong/16873/2016, DK16873) isolated from southern China in 2016 could replicate in multiple organs of inoculated ducks. DK16873 virus caused mild infections and killed 2/5 of inoculated ducks, and GS16568 virus did not kill inoculated ducks. In addition, the two viruses could be transmitted via direct contact between ducks. DK16873 and GS16568 viruses killed 2/5 and 1/5 of contact ducks, respectively. Furthermore, ducks inoculated with the two H5N6 viruses exhibited different expressions of immune-related genes in their lungs. The expression of RIG-I, TLR3 and IL6 was significantly upregulated at 12 h post-inoculation (HPI) and most of the tested immune-related genes were significantly upregulated at 3 days post-inoculation (DPI). Notably, the expression of RIG-I and IL-6 in response to DK16873 virus was significantly higher than for GS16568 virus at 12 HPI and 3 DPI. Our research have provided helpful information about the pathogenicity, transmission and immune-related genes expression in ducks infected with new H5N6 AIVs. [ABSTRACT FROM AUTHOR]

Grass carp ( Ctenopharyngodon idellus ) is an economical aquaculture species in China, and the Grass Carp Reovirus (GCRV) that causes hemorrhagic disease seriously affects the grass carp cultivation industry. Substantial evidence indicates that there is an association between the membrane-associated RING-CH family of E3 ligase (MARCH) family and immune defense in mammals, while functional studies on non-mammalian MARCH proteins are limited. In order to know the characteristics of the MARCH genes in C. idellus , eight MARCH genes ( MARCH1, 2, 5, 6, 7, 8, 9 and 11 ) were cloned and the open reading frames (ORF) were identified in grass carp. All MARCH proteins in grass carp contained an RING-CH domain, which is characteristic of the MARCH protein. The phylogenetic analysis revealed that different MARCH proteins gathered into their separate clusters. All eight members of the MARCH gene family were detected in all tissues sampled, but the relative expression level differed. In addition, the mRNA expression of all the MARCHs was regulated at different levels in the immune organs after a GCRV challenge, and they responded robustly in both the intestine and liver. The mRNA expression of MARCH8, MHC II, TfR, IL1RAP, EGR1, and DUSP1 in the intestine after GCRV infection was analyzed, and the results showed that MARCH8 could negatively regulate TfR, IL1RAP, EGR1, and DUSP1, which signaled via the MAPK or NF-κB-activation pathways that play vital roles in immunity. Our findings identified a novel gene family in C. idellus and provided novel evidence that MARCH genes are inducible and involved in the immune response. Moreover, MARCH8 might function to negatively regulate immune receptors in C. idellus . Therefore, the MARCH might play a vital role in regulating the immune response of C. idellus . [ABSTRACT FROM AUTHOR]

Abstract: Sea cucumber (Apostichopus japonicus) is one of the most important aquaculture species in China. To profile gene expression in different tissues and to identify immune-related genes, we constructed three independent cDNA libraries that were derived from intestine (Int), body wall (BW) and respiratory tree (RT) of healthy individuals. From these libraries, a total of 5728 high-quality expressed sequence tags (ESTs) of longer than 100bp were generated. Clustering and assembly of the ESTs identified 495 contigs and 2118 singletons, representing 2613 unigenes. Basic Local Alignment Search Tool (BLAST) analysis showed that 902 unigenes (34.52%) had significant homology (E value ≤10−5) to genes with known or putative functions in GenBank. These unigenes were grouped into 18 functional categories. When compared among these three libraries, RT library showed fewer categories (9) while the other two libraries displayed more (17 and 16 for Int and BW libraries respectively). Considering the components among the three cDNA libraries, 655, 259 and 484 tissue-specific unigenes were observed in Int, BW and RT libraries respectively; however, only 95 unigenes were common to all three libraries. The most abundant contigs in the three libraries were identified as genes related to cell and organism defense (551 ESTs), metabolism (415 ESTs), translation/ribosomal structure and biogenesis (342 ESTs), gene/protein expression (178 ESTs), structure and motility (75 ESTs), energy production and conversion (51 ESTs), and cytoskeleton (39 ESTs). The list of 636 ESTs that matched with immune-related genes were categorized as pattern recognition receptors and signaling, complement components, transferrin superfamily members, cytokines and growth factors, and effector genes. The present manuscript presents a fair amount of data regarding the spatial expression pattern of many genes (BW vs. Int vs. RT of A. japonicus). [Copyright &y& Elsevier]

The interaction between a host and its parasitoid is one of the most fascinating relationships of insects. Immune-related genes play crucial roles in this association. Nevertheless, until now, identification of these genes on a large scale has not received much attention. To gain insight into the parasitic effects of the endoparasitoid Aulacocentrum confusum (Hymenoptera: Braconidae) on Glyphodes pyloalis (Lepidoptera: Pyralidae) larva, which is a destructive pest of mulberry (Morus alba L.) trees in China, we presented a transcriptome dataset for uncovering immune-related genes in parasitized G. pyloalis larvae. In total, 91,118,138 and 92,778,814 clean reads were obtained from parasitized and healthy host larvae, respectively, and de novo assembly generated 57,122 unigenes. The transcriptional profile of G. pyloalis larvae was remarkably influenced by parasitism. A total of 3259 differentially expressed genes (DEGs) were identified in parasitized and nonparasitized G. pyloalis larvae and 55 genes related to immune response were screened from these DEGs. Among the 55 DEGs, 37 genes were significantly upregulated, and 18 genes were downregulated. qRT-PCR validated the sequencing results and revealed that the expression levels of selected immune-related genes depended on the parasitization and duration after parasitization. Knocking down the C type lectin gene (CTL) changed the expression of serine proteinase , serine protease inhibitor , antimicrobial peptide , prophenoloxidase activating enzymes and peroxiredoxin in G. pyloalis larvae, suggesting CTL can modulate the immune response after parasitization by A. confusum females. The present study provides a foundation for revealing the molecular mechanisms of immune response in G. pyloalis larvae when they are parasitized by A. confusum and promotes the development of novel biological control practices for G. pyloalis. [Display omitted] • A transcriptome sequencing of Glyphodes pyloalis larvae was performed. • A total of 55 immune-related genes in respond to parasitization were identified. • The expression of selected immune-related genes was variable after parasitization. • C type lectin can modulate the expression of genes in the immune pathway. [ABSTRACT FROM AUTHOR]

Hucho bleekeri is a glacial relict and critically endangered fish restricted to the Yangtze River drainage in China. The lack of basic genomic information and immune characteristics will hinder the way toward protecting this species. In the present study, we conducted the first transcriptome analysis of H. bleekeri using the combination of SMRT and Illumina sequencing technology. Transcriptome sequencing generated a total of 93,330 non-redundant full-length unigenes with a mean length of 3072 bp. A total of 92,472 (99.08%) unigenes were annotated in at least one of the Nr protein, Swiss-Prot, KEGG, KOG, GO, Nt and Pfam databases. KEGG analysis showed that a total of 7240 unigenes belonging to 28 immune pathways were annotated to the immune system category. Meanwhile, differentially expressed genes between mucosa-associated tissues (skin, gill and hindgut) and systemic-immune tissues (spleen, head kidney and liver) were obtained. Importantly, genes participating in diverse immune signalling pathways and their expression profiles in H. bleekeri were discussed. In addition, a large number of long non-coding RNAs (lncRNAs) and simple sequence repeats (SSRs) were obtained in the H. bleekeri transcriptome. The present study will provide basic genomic information for H. bleekeri and for further research on analysing the characteristics of both the innate and adaptive immune systems of this critically endangered species. • The first transcriptome sequencing of Hucho bleekeri was obtained using SMRT and Illumina technology. • A large number of full-length unigenes were obtained and successfully annotated. • Unigenes involved in the immune system and DEGs between mucosa-associated tissues and systemic-immune tissues were obtained. • Genes participating in diverse immune signalling pathways and their expression profiles in H. bleekeri were discussed. [ABSTRACT FROM AUTHOR]